Functional analysis of ERM proteins in epithelial morphogenesis

ERM蛋白在上皮形态发生中的功能分析

• 批准号: 7783553
• 负责人: Richard G Fehon
• 金额: $ 50.5万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7783553
• 关键词: Actins; Adherens Junction; Adhesions; Apical; Biochemistry; Biological; Biological Models; Biological Process; Cell Shape; Cell membrane; Cell physiology; Cells; Cellular biology; Chicago; Complex; Cytoplasmic Protein; Cytoskeleton; Disease; Drosophila genus; Endocytosis; Epithelial; Epithelial Cells; Epithelium; Family; Genetic; Genetic Models; Goals; Homeostasis; Integral Membrane Protein; Intercellular Junctions; Intestines; Laboratories; Link; Mammalian Cell; Mammals; Mediating; Membrane; Modeling; Molecular; Morphogenesis; Mus; Neoplasm Metastasis; Output; Phenotype; Phospholipids; Phosphorylation; Phosphotransferases; Play; Post-Translational Protein Processing; Process; Property; Proteins; Proteomics; Receptor Signaling; Regulation; Research; Research Personnel; Role; Side; Signal Transduction; Sterility; Surface; System; Testing; Tissues; Universities; Work; design; ezrin; fly; in vivo; insight; intercellular communication; intestinal epithelium; moesin; mutant; novel; polarized cell; protein complex; public health relevance; radixin protein; research study; response; rho; segregation; tool

DESCRIPTION (provided by applicant): The ability to form specialized membrane domains composed of unique sets of transmembrane proteins, associated cytoplasmic proteins, and phospholipids, is a fundamental property of polarized epithelial cells. Membrane domains, such as the apical surface or junctional complex, allow spatial segregation of functions at the plasma membrane that are essential for polarized epithelia. Central to this process is the formation of protein complexes on the cytoplasmic side of the membrane that localize transmembrane proteins, regulate their signaling output and control their abundance via regulated endocytosis. The Ezrin, Radixin, Moesin (ERM) proteins organize a key role in this process. In this proposal we describe experiments designed to take advantage of the combined expertise of two investigators, Andrea McClatchey (Harvard/MGH) and Richard Fehon (University of Chicago), to extend our understanding of ERM function. The investigators and their laboratories bring together expertise in two powerful experimental systems, the mouse and Drosophila. The proposed research utilizes a multifaceted approach, including genetics, biochemistry, cell biology and proteomics to better understand the functions of these highly conserved proteins. Specifically, we plan to: 1) Determine the molecular mechanisms that link the ERM proteins to the activation state of Rho in developing epithelial cells. 2) Examine the molecular mechanisms that regulate ERM activity, particularly in the context of how ERM activity is dynamically regulated. 3) Delineate the function of the ERM proteins in cell:cell junction remodeling. 4) To build an integrated model of ERM-mediated complex formation. These experiments are expected to provide novel insight into the functions of ERM proteins in biological processes such as apical-basal polarity cytoskeletal regulation, intestinal lumen formation and homeostasis, and metastasis. They should also yield a better understanding of the cellular processes that establish specialized membrane domains in polarized cells, and inform our understanding of cytoskeletal and junctional dynamics during morphogenesis and in disease. PUBLIC HEALTH RELEVANCE: We will carry out complementary and synergistic studies in two powerful model systems - Drosophila and the mouse - to examine the function of ERM proteins in epithelial morphogenesis. Specifically, we will test the hypothesis that ERM proteins provide local regulation of RhoGTPase activity in response to upstream signals/receptors using molecular tools and genetic models that we have developed in each system. We will also delineate the function of ERM proteins in cell junction remodeling/stability as suggested by the phenotypes of ERM loss in both flies and mammals.

描述（由申请人提供）：形成由独特的跨膜蛋白组、相关细胞质蛋白和磷脂组成的专门膜域的能力是极化上皮细胞的基本特性。膜域，例如顶端表面或连接复合体，允许质膜上的功能空间分离，这对于极化上皮至关重要。该过程的核心是在膜的细胞质侧形成蛋白质复合物，该复合物定位跨膜蛋白质，调节其信号输出并通过调节的内吞作用控制其丰度。 Ezrin、Radixin、Moesin (ERM) 蛋白在此过程中发挥着关键作用。 在本提案中，我们描述了旨在利用两位研究人员 Andrea McClatchey（哈佛大学/麻省总医院）和 Richard Fehon（芝加哥大学）的综合专业知识的实验，以扩展我们对 ERM 功能的理解。研究人员及其实验室汇集了小鼠和果蝇这两个强大实验系统的专业知识。拟议的研究利用多方面的方法，包括遗传学、生物化学、细胞生物学和蛋白质组学，以更好地了解这些高度保守的蛋白质的功能。具体来说，我们计划： 1) 确定在发育中的上皮细胞中将 ERM 蛋白与 Rho 激活状态联系起来的分子机制。 2) 检查调节 ERM 活性的分子机制，特别是在如何动态调节 ERM 活性的背景下。 3）描述ERM蛋白在细胞：细胞连接重塑中的功能。 4）建立ERM介导的复合物形成的综合模型。这些实验有望为 ERM 蛋白在生物过程中的功能提供新的见解，例如顶端-基底极性细胞骨架调节、肠腔形成和稳态以及转移。它们还应该更好地理解在极化细胞中建立专门的膜域的细胞过程，并帮助我们理解形态发生和疾病期间的细胞骨架和连接动态。 公共健康相关性：我们将在两个强大的模型系统（果蝇和小鼠）中进行互补和协同研究，以检查 ERM 蛋白在上皮形态发生中的功能。具体来说，我们将使用我们在每个系统中开发的分子工具和遗传模型来测试 ERM 蛋白响应上游信号/受体而提供 RhoGTPase 活性的局部调节的假设。我们还将描述 ERM 蛋白在细胞连接重塑/稳定性中的功能，正如果蝇和哺乳动物中 ERM 丧失的表型所表明的那样。